GSM6934372: Cross-contamination experiment: Liver, mouse 4, replicate 3; Mus musculus; OTHER
The amplified cDNA was purified using SPRIselectTM beads (Beckman Coulter) following a left sided size selection with a bead-to-sample ratio of 0.8X. In brief, 160 µL of sample were mixed with 128 µL of resuspended SPRIselect beads and incubated for 5 min at rt. Beads were washed two times in 85% ethanol and air-dried for 3 min. The cDNA was eluted in 20 µL ultrapure water by incubation at 37 °C for 10 min. The supernatants were transferred to a fresh tube resulting in 9 tubes of purified cDNA. The quality of the cDNA was analyzed using the Bioanalyzer High Sensitivity DNA chip (Agilent) and samples were stored at -20 °C. The concentration of the cDNA was determined using a Qubit 1X dsDNA assay (Invitrogen) and the sequencing library was generated using the Nextera XT DNA Library Preparation Kit (Illumina). The quality of the library was assessed using the Bioanalyzer High Sensitivity DNA chip (Agilent). Samples were sequenced on a NovaSeq 6000 system (Illumina) at a sequencing depth of minimum 50,000 reads per spot using a 100 cycles kit in paired-end mode. Following read length configurations were used: R1: 7 cycles, i7: 6 cycles, R2: 58 cycles.